Phytochemical constituents of pressurized liquid extract from Ziziphus jujubа Mill. (Rhamnaceae) fruits and in vitro inhibitory activity on α-glucosidase, pancreatic α-amylase and lipase

The phytochemical composition of extract from Ziziphus jujubа Mill (Rhamnaceae) fruits obtained by pressurized liquid extraction (PLE) was investigated in the current study. The phytochemical analysis showed the presence of triterpenes, flavonoids and phenolic acids. The fruit extract had a total phenolic content of 21 ±2.01 mg gallic acid equivalents per g of extract. (mg GAE/g extract), and total flavonoid content 2.48 ± 0.24 mg catechin equivalents per g dried extract (mg CE/g extract). This study evaluated the in vitro antioxidant activity of jujube extract and its inhibitory effects on α-glucosidase and pancreatic α-amylase and lipase. The obtained results proved inhibition of α-glucosidase with 70.94% and inhibition of α-amylase with 56.08%. Enzyme kinetic studies indicated that the inhibitors in the extract have the most potent inhibition of lipase compared to the other two enzymes with inhibitory constant Ki of 0.074 mg/cm3 . These results suggest the possible application of extract from Z. jujubа fruits obtained by PLE in the management of metabolic disorders as a whole. These are the first studies on this plant species cultivated in Bulgari

Tailoring tobacco hairy root metabolism for the production of stilbenes

Tobacco hairy root (HR) cultures, which have been widely used for the heterologous production of target compounds, have an innate capacity to bioconvert exogenous t-resveratrol (t-R) into t-piceatannol (t-Pn) and t-pterostilbene (t-Pt). We established genetically engineered HR carrying the gene encoding stilbene synthase (STS) from Vitis vinifera and/or the transcription factor (TF) AtMYB12 from Arabidopsis thaliana, in order to generate a holistic response in the phenylpropanoid pathway and coordinate the up-regulation of multiple metabolic steps. Additionally, an artificial microRNA for chalcone synthase (amiRNA CHS) was utilized to arrest the normal flux through the endogenous chalcone synthase (CHS) enzyme, which would otherwise compete for precursors with the STS enzyme imported for the flux deviation. The transgenic HR were able to biosynthesize the target stilbenes, achieving a production of 40 μg L-1 of t-R, which was partially metabolized into t-Pn and t-Pt (up to 2.2 μg L-1 and 86.4 μg L-1, respectively), as well as its glucoside piceid (up to 339.7 μg L-1). Major metabolic perturbations were caused by the TF AtMYB12, affecting both primary and secondary metabolism, which confirms the complexity of biotechnological systems based on seed plant in vitro cultures for the heterologous production of high-value molecules

Tailoring tobacco hairy root metabolism for the production of stilbenes

Tobacco hairy root (HR) cultures, which have been widely used for the heterologous production of target compounds, have an innate capacity to bioconvert exogenous t-resveratrol (t-R) into t-piceatannol (t-Pn) and t-pterostilbene (t-Pt). We established genetically engineered HR carrying the gene encoding stilbene synthase (STS) from Vitis vinifera and/or the transcription factor (TF) AtMYB12 from Arabidopsis thaliana, in order to generate a holistic response in the phenylpropanoid pathway and coordinate the up-regulation of multiple metabolic steps. Additionally, an artificial microRNA for chalcone synthase (amiRNA CHS) was utilized to arrest the normal flux through the endogenous chalcone synthase (CHS) enzyme, which would otherwise compete for precursors with the STS enzyme imported for the flux deviation. The transgenic HR were able to biosynthesize the target stilbenes, achieving a production of 40 μg L−1 of t-R, which was partially metabolized into t-Pn and t-Pt (up to 2.2 μg L−1 and 86.4 μg L−1, respectively), as well as its glucoside piceid (up to 339.7 μg L−1). Major metabolic perturbations were caused by the TF AtMYB12, affecting both primary and secondary metabolism, which confirms the complexity of biotechnological systems based on seed plant in vitro cultures for the heterologous production of high-value molecules.This work has been supported by a grant from the Spanish Ministry of Science and Innovation (BIO2014-51861-R, BIO2017-82374-R), Generalitat de Catalunya (2014SGR215). Diego Hidalgo is a predoctoral fellow of Mexican CONACyT

Plant In Vitro Systems as a Sustainable Source of Active Ingredients for Cosmeceutical Application

Cosmeceuticals are hybrids between cosmetics and pharmaceuticals which are being designed for a dual purpose: (1) To provide desired esthetical effects and (2) simultaneously treat dermatological conditions. The increased demand for natural remedies and the trends to use natural and safe ingredients resulted in intensive cultivation of medicinal plants. However, in many cases the whole process of plant cultivation, complex extraction procedure, and purification of the targeted molecules are not economically feasible. Therefore, the desired production of natural cosmetic products in sustainable and controllable fashion in the last years led to the intensive utilization of plant cell culture technology. The present review aims to highlight examples of biosynthesis of active ingredients derived through plant in vitro systems with potential cosmeceutical application. The exploitation of different type of extracts used in a possible cosmeceutical formulation, as well as, their activity tested in in vitro/in vivo models is thoroughly discussed. Furthermore, opportunities to manipulate the biosynthetic pathway, hence engineering the biosynthesis of some secondary metabolites, such as anthocyanins, have been highlighted

Cultivation of diploid and tetraploid hairy roots of Datura stramonium L. in stirred tank bioreactor for tropane alkaloids production

Biomass accumulation and tropane alkaloids production by diploid and tetraploid hairy root cultures of Datura stramonium L. cultivated in stirred tank bioreactor at different aeration rates were investigated. The maximal growth for both hairy root cultures (ADB = 8.3 g/L and 6.8 g/L for diploid and tetraploid line, respectively) was achieved at aeration rate of 15.0 L/(L.h). The corresponding growth indexes were remarkably high (GIDW = 9.0 and 7.8 for diploid and tetraploid line, respectively) compared to the values, usually reported for other hairy root cultures. The optimal aeration rate for biomass accumulation was also optimal for alkaloids biosynthesis. According to our survey, the achieved maximal amounts of accumulated hyoscyamine (35.0 mg/L and 27.0 mg/L for diploid and tetraploid line) were the highest reported in the scientific literature for D. stramonium L. hairy roots. During the cultivation in stirred tank bioreactor, the hairy roots biosynthesized pharmaceutically important alkaloid scopolamine in minor concentrations. This is an important observation since scopolamine was not detected during submerged cultivation of these hairy root lines in other bioreactor types. However, the ploidy level was found to be the most important factor concerning scopolamine production by D. stramonium L. hairy roots cultures. The present work demonstrated the effect of ploidity levels on biomass accumulation and tropane alkaloids production by D. stramonium L. hairy roots cultivated in stirred tank bioreactor. This investigation show that the stirred tank bioreactor could be successfully applied for both maximal biomass accumulations, as well as for manipulation of tropane alkaloids production by diploid and tetraploid D. stramonium L. hairy root cultures

<i>Crocus sativus</i> Extract as a Biological Agent for Disease-Modifying Therapy of Collagenase-Induced Mouse Model of Osteoarthritis

Objectives: Osteoarthritis (OA) is an age-related joint disease that involves the degeneration of cartilage and is the most prevalent form of arthritis, affecting a large part of the population. OA is a multifactorial disorder, and no single etiological mechanism has been found to be common to all forms of the disease. Currently used therapies for control of the disease are mainly nonsteroidal anti-inflammatory drugs (NSAIDs) and corticosteroid medications. The aim of this study was to investigate the extract from Crocus sativus as a biological disease-suppressing therapy agent. Methods: Balb/c mice were injected intra-articularly with Clostridium histolyticum type IA for induction of osteoarthritis. The mice were randomized to five groups: control group, I group (CIOA untreated), II group (CIOA + 100 mg/kg/daily saffron), III group (CIOA + 50 mg/kg/daily saffron), IV group (CIOA + 25 mg/kg/daily saffron). Flow-cytometry analysis was used to study the splenocytes’ phenotype isolated from the treated animals. The serum levels of inflammatory and anti-inflammatory cytokines were analyzed with ELISA. The histological assessment was used to analyze the saffron extract effect on histopathological alterations. Results: Saffron treatment significantly decreased osteoarthritis-associated joint histological manifestations and decreased serum TNFα levels. The flow-cytometry analysis showed a decrease in pro-inflammatory immune cell subtypes in the spleen. Conclusions: The results obtained suggest that saffron affected the disease progression and could be a potential therapeutic approach in osteoarthritic patients’ therapy

Acetylcholinesterase inhibitory, antioxidant, and antimicrobial activities of Salvia tomentosa Mill. essential oil

Chemical composition and bioactivity of essential oil from Salvia tomentosa Mill. natively grown in Bulgaria were investigated. GC-MS analysis identified 60 compounds which represented 98% of the oil constituents. The prevalent constituents were monoterpenes with eight dominant compounds being identified: borneol (10.3%), β-pinene (9%), camphor (7.9%), α-pinene (6%), camphene (4%), 1.8-cineole (3.8%), α-limonene (3.5%) and β-caryophyllene (3%). The essential oil showed considerable acetylcholinesterase inhibitory activity (IC50=0.28±0.06 µg/mL), comparable with that of galanthamine. Study of antioxidant activity strongly suggested that the hydrogen atom transfer reaction was preferable over the electron transfer (ORAC=175.0±0.40 µM Trolox equivalents/g oil and FRAP=1.45±0.21 mM Trolox equivalents/g oil). The essential oil showed moderate antifungal and antibacterial activities against Candida albicans and Gram-positive bacteria, whereas it was almost inactive against the investigated Gram-negative strains. The results suggested that the essential oil of Bulgarian S. tomentosa could be considered as a prospective active ingredient for prevention of oxidative stress-related and neurodegenerative disorders in aromatherapy. Because of the high antioxidant capacity, the oil could be considered as natural supplement or antioxidant in cosmetics and food products

Biotechnologically-Produced Myconoside and Calceolarioside E Induce Nrf2 Expression in Neutrophils

The pathological manifestation of various diseases can be suppressed by the activation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2), a transcriptional regulator of the cellular redox balance. Haberlea rhodopensis Friv. is a resurrection plant species endemic for Bulgaria, containing biologically active phenylethanoid glycosides that might possess antioxidant or redox activity. This study aimed to analyze the metabolic profile of in vitro cultured H. rhodopensis and to identify molecules that increase Nrf2 expression in bone marrow neutrophils. Fractions B, D, and E containing myconoside, or myconoside and calceolarioside E in ratios 1:0.6 and 0.25:1 were found to be the most active ones. Fraction B (200 µg/mL) improved neutrophil survival and strongly increased the Nrf2 intracellular level, while D and E, as well as, myconoside and calceolarioside E at the same ratios had a superior effect. Calceolarioside E (32 µg/mL) had stronger activity than myconoside, the effect of which was very similar to that of 2-cyano-3,12-dioxo-oleana-1,9(11)-dien-28-oic acid methyl ester (CDDO-Me), used as a positive control. These data indicate that both molecules, used alone or in combination have stimulatory activity on the endogenous Nrf2 level, indicating their therapeutic potential to regulate the cellular redox homeostasis oxidative stress-associated pathologies